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Article: Serological diagnosis of nasopharyngeal carcinoma by enzyme linked immunosorbant assay: Optimization, standardization and diagnostic criteria

TitleSerological diagnosis of nasopharyngeal carcinoma by enzyme linked immunosorbant assay: Optimization, standardization and diagnostic criteria
Authors
Issue Date1998
PublisherZhonghua Yixuehui. The Journal's web site is located at http://www.cmj.org/
Citation
Chinese Medical Journal, 1998, v. 111 n. 6, p. 531-536 How to Cite?
AbstractObjectives To produce an enzyme linked immunosorbant assay (ELISA) to detect antibodies against Epstein Barr virus (EBV) specified nuclear antigen 1 (EBNA 1) and the 18kD EBV matrix protein, and to determine and optimize its sensitivity and specificity for the diagnosis of nasopharyngeal carcinoma (NPC). Methods We used a combination of highly purified glutathione transferase fusion proteins of the 40kD carboxy domain of EBNA1 and the 18kD EBV matrix protein for coating ELISA plates. In three separate studies, we tested for IgA antibodies in serum specimens from 28 EBV seronegative donors, 284 EBV seropositive donors and 160 newly diagnosed NPC patients. By comparing the sensitivity and specificity of diagnosis obtained for different cutoff values, we derived several quantitative parameters to evaluate assay performance, establish objective diagnostic criteria which optimize the intrinsic diagnostic capability of the assay and assess the significance of individual test results, respectively. Optimum cutoff optical density (OD) is defined as the cutoff OD where sensitivity of the assay equals its specificity, and resolution of the assay is indicated by the value of sensitivity (or specificity) determined at the optimum cutoff OD. Diagnosis of NPC was achieved by setting a cutoff zone at + / 20% of this value. Results All the EBV seronegative donors tested were not reactive, and most of the EBV seropositive donors were weakly reactive, while the majority of NPC patients were moderately or strongly reactive. While the assay was thus shown to be specific for EBV, there was an overlap in the level of these serum antibodies between few individuals of the two latter groups. It was shown that the assay performed equally well in two separate studies conducted under different testing conditions and using different collections of sera in that assay resolution determined on these occasions were 86% and 87% respectively. Diagnosis of NPC can be achieved at the same expected sensitivity of 89% and 83% determined at the lower and upper limits of the cutoff zones, with the corresponding values of specificity being 78% and 91%. It was further shown in the third study that resolution of the assay can be increased to 90% using an assay produced with a higher concentration of the same antigens, and that diagnosis of NPC can be achieved at a higher sensitivity ranging between 86% and 95% at a corresponding specificity of 93% and 86%. Conclusions After optimization and standardization, the ELISA can achieve a sensitivity ranging from 86% to 95%, with corresponding specificities of 93% and 86% respectively for the diagnosis of NPC.
Persistent Identifierhttp://hdl.handle.net/10722/78951
ISSN
2015 Impact Factor: 0.957
2015 SCImago Journal Rankings: 0.428
References

 

DC FieldValueLanguage
dc.contributor.authorNg, MHen_HK
dc.contributor.authorChen, HLen_HK
dc.contributor.authorLuo, RXen_HK
dc.contributor.authorChan, KHen_HK
dc.contributor.authorWoo, PCYen_HK
dc.contributor.authorSham, JSTen_HK
dc.contributor.authorHuang, Jen_HK
dc.contributor.authorSeto, WHen_HK
dc.contributor.authorSmith, Pen_HK
dc.contributor.authorGriffin, BEen_HK
dc.date.accessioned2010-09-06T07:48:47Z-
dc.date.available2010-09-06T07:48:47Z-
dc.date.issued1998en_HK
dc.identifier.citationChinese Medical Journal, 1998, v. 111 n. 6, p. 531-536en_HK
dc.identifier.issn0366-6999en_HK
dc.identifier.urihttp://hdl.handle.net/10722/78951-
dc.description.abstractObjectives To produce an enzyme linked immunosorbant assay (ELISA) to detect antibodies against Epstein Barr virus (EBV) specified nuclear antigen 1 (EBNA 1) and the 18kD EBV matrix protein, and to determine and optimize its sensitivity and specificity for the diagnosis of nasopharyngeal carcinoma (NPC). Methods We used a combination of highly purified glutathione transferase fusion proteins of the 40kD carboxy domain of EBNA1 and the 18kD EBV matrix protein for coating ELISA plates. In three separate studies, we tested for IgA antibodies in serum specimens from 28 EBV seronegative donors, 284 EBV seropositive donors and 160 newly diagnosed NPC patients. By comparing the sensitivity and specificity of diagnosis obtained for different cutoff values, we derived several quantitative parameters to evaluate assay performance, establish objective diagnostic criteria which optimize the intrinsic diagnostic capability of the assay and assess the significance of individual test results, respectively. Optimum cutoff optical density (OD) is defined as the cutoff OD where sensitivity of the assay equals its specificity, and resolution of the assay is indicated by the value of sensitivity (or specificity) determined at the optimum cutoff OD. Diagnosis of NPC was achieved by setting a cutoff zone at + / 20% of this value. Results All the EBV seronegative donors tested were not reactive, and most of the EBV seropositive donors were weakly reactive, while the majority of NPC patients were moderately or strongly reactive. While the assay was thus shown to be specific for EBV, there was an overlap in the level of these serum antibodies between few individuals of the two latter groups. It was shown that the assay performed equally well in two separate studies conducted under different testing conditions and using different collections of sera in that assay resolution determined on these occasions were 86% and 87% respectively. Diagnosis of NPC can be achieved at the same expected sensitivity of 89% and 83% determined at the lower and upper limits of the cutoff zones, with the corresponding values of specificity being 78% and 91%. It was further shown in the third study that resolution of the assay can be increased to 90% using an assay produced with a higher concentration of the same antigens, and that diagnosis of NPC can be achieved at a higher sensitivity ranging between 86% and 95% at a corresponding specificity of 93% and 86%. Conclusions After optimization and standardization, the ELISA can achieve a sensitivity ranging from 86% to 95%, with corresponding specificities of 93% and 86% respectively for the diagnosis of NPC.en_HK
dc.languageengen_HK
dc.publisherZhonghua Yixuehui. The Journal's web site is located at http://www.cmj.org/en_HK
dc.relation.ispartofChinese Medical Journalen_HK
dc.titleSerological diagnosis of nasopharyngeal carcinoma by enzyme linked immunosorbant assay: Optimization, standardization and diagnostic criteriaen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0366-6999&volume=111&issue=6&spage=531&epage=536&date=1998&atitle=Serological+diagnosis+of+nasopharyngeal+carcinoma+by+enzyme+linked+immunosorbant+assay:+optimization,+standardization+and+diagnostic+criteriaen_HK
dc.identifier.emailChen, HL:hlchen@hkucc.hku.hken_HK
dc.identifier.emailWoo, PCY:pcywoo@hkucc.hku.hken_HK
dc.identifier.authorityChen, HL=rp00383en_HK
dc.identifier.authorityWoo, PCY=rp00430en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.scopuseid_2-s2.0-0032081855en_HK
dc.identifier.hkuros36680en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0032081855&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume111en_HK
dc.identifier.issue6en_HK
dc.identifier.spage531en_HK
dc.identifier.epage536en_HK
dc.publisher.placeChinaen_HK
dc.identifier.scopusauthoridNg, MH=7202076421en_HK
dc.identifier.scopusauthoridChen, HL=26643315400en_HK
dc.identifier.scopusauthoridLuo, RX=7202671104en_HK
dc.identifier.scopusauthoridChan, KH=35338760600en_HK
dc.identifier.scopusauthoridWoo, PCY=7201801340en_HK
dc.identifier.scopusauthoridSham, JST=24472255400en_HK
dc.identifier.scopusauthoridHuang, J=7407189939en_HK
dc.identifier.scopusauthoridSeto, WH=7005799377en_HK
dc.identifier.scopusauthoridSmith, P=7406997818en_HK
dc.identifier.scopusauthoridGriffin, BE=7201402684en_HK

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